Required Practical 8 Flashcards
comparing electron transfer in mitochondria and chloroplasts
the seed coat of each bean was removed and discarded.
suggest why?
better release of mitochondria
what is meant by isotonic?
same water potential as variable e.g. in this case MITOCHONDRIA
why were the beans blended?
release mitochondria from cells
the cloth was moistened with isolation medium and not moistened with water.
explain why water was not used to moisten the cloth?
- water would affect WP
- as has higher WP
- water would enter mitochondria by OSMOSIS
the mixture was poured through cloth to filter it.
give 2 reasons why it was necessary to filter the mixture?
- to remove cell debris
- remove large organelles
why was the mitochondrial suspension kept on ice before use?
reduce enzyme activity
succinate was added to the suspension of mitochondria as a respiratory substrate.
explain why glucose could NOT be used as the respiratory substrate in this investigation?
- glucose too big to cross mitochondrial membrane
- no cytoplasm
- so no glycolysis
in tube A were the following:
- 5 cm3 succinate sol
- 2cm3 mitochondrial suspension
- 10 drops DCPIP
the contents of tube A did not go colourless.
suggest one reason why?
mitochondrial suspension still present
addition of succinate to mitochondria leads to electron transfer.
what leads to electron transfer in chloroplast investigation?
explain your answer?
- light
- light excites chlorophyll
how much chloroplast suspension, DCPIP, distilled water and isolation medium would you put in a control tube?
- 0 cm3 chloroplast suspension
- 5cm3 DCPIP
- 1cm3 distilled water
- 1cm3 isolation medium
you were told not to put the leaf midrib or leaf stalk in blender.
why?
contain less chloroplasts
what is the purpose of using a blender?
break open organelles
the isolation medium was cold and was also isotonic with the spinach tissue.
explain why it was important that the isolation medium was cold?
- reduce enzyme activity
- enzymes released during blending
- chloroplast not damaged
explain why it was important that the isolation medium was isotonic with the spinach tissue?
- so WP same as plant tissue
- chloroplasts dont burst
there are large pieces of tissue and other organelles in your chloroplast suspension.
describe how you could isolate the chloroplasts from the other components in the suspension?
- filter using centrifuge
- at low speed then high speed
- collect pellets